Article

Reprogramming of mouse and human somatic cells by high-performance engineered factors.

The State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 320 Yue-Yang Road, Shanghai 200031, China.
EMBO Reports (impact factor: 7.36). 03/2011; 12(4):373-8. DOI:10.1038/embor.2011.11 pp.373-8
Source: PubMed

ABSTRACT Reprogramming somatic cells to become induced pluripotent stem cells (iPSCs) by using defined factors represents an important breakthrough in biology and medicine, yet remains inefficient and poorly understood. We therefore devised synthetic factors by fusing the VP16 transactivation domain to OCT4 (also known as Pou5f1), NANOG and SOX2, respectively. These synthetic factors could reprogramme both mouse and human fibroblasts with enhanced efficiency and accelerated kinetics. Remarkably, Oct4-VP16 alone could efficiently reprogramme mouse embryonic fibroblasts (MEFs) into germline-competent iPSCs. Furthermore, episomally delivered synthetic factors could reproducibly generate integration-free iPSCs from MEFs with enhanced efficiency. Our results not only demonstrate the feasibility of engineering more potent reprogramming factors, but also suggest that transcriptional reactivation of OCT4 target genes might be a rate-limiting step in the conversion of somatic cells to pluripotent cells. Synthetic factor-based reprogramming might lead to a paradigm shift in reprogramming research.

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    Article: Generation of mouse induced pluripotent stem cells without viral vectors.
    Keisuke Okita, Masato Nakagawa, Hong Hyenjong, Tomoko Ichisaka, Shinya Yamanaka
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    ABSTRACT: Induced pluripotent stem (iPS) cells have been generated from mouse and human somatic cells by introducing Oct3/4 and Sox2 with either Klf4 and c-Myc or Nanog and Lin28 using retroviruses or lentiviruses. Patient-specific iPS cells could be useful in drug discovery and regenerative medicine. However, viral integration into the host genome increases the risk of tumorigenicity. Here, we report the generation of mouse iPS cells without viral vectors. Repeated transfection of two expression plasmids, one containing the complementary DNAs (cDNAs) of Oct3/4, Sox2, and Klf4 and the other containing the c-Myc cDNA, into mouse embryonic fibroblasts resulted in iPS cells without evidence of plasmid integration, which produced teratomas when transplanted into mice and contributed to adult chimeras. The production of virus-free iPS cells, albeit from embryonic fibroblasts, addresses a critical safety concern for potential use of iPS cells in regenerative medicine.
    Science 11/2008; 322(5903):949-53. · 31.20 Impact Factor
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Keywords

factors
 
fusing
 
germline-competent iPSCs
 
human fibroblasts
 
induced pluripotent
 
integration-free iPSCs
 
NANOG
 
OCT4
 
OCT4 target genes
 
pluripotent cells
 
potent reprogramming factors
 
rate-limiting step
 
reprogramming research
 
Reprogramming somatic cells
 
Synthetic factor-based reprogramming
 
synthetic factors
 
transcriptional reactivation
 
VP16 transactivation domain